title: Modulation of chaperone gene expression in mutagenized Saccharomyces cerevisiae strains developed for recombinant human albumin production results in increased production of multiple heterologous proteins.
abstract: The yeast Saccharomyces cerevisiae has been successfully established as a commercially viable system for the production of recombinant proteins. Manipulation of chaperone gene expression has been utilized extensively to increase recombinant protein production from S. cerevisiae, focusing predominantly on the products of the protein disulfide isomerase gene PDI1 and the hsp70 gene KAR2. Here we show that the expression of the genes SIL1, LHS1, JEM1, and SCJ1, all of which are involved in regulating the ATPase cycle of Kar2p, is increased in a proprietary yeast strain, developed by several rounds of random mutagenesis and screening for increased production of recombinant human albumin (rHA). To establish whether this expression contributes to the enhanced-production phenotype, these genes were overexpressed both individually and in combination. The resultant strains showed significantly increased shake-flask production levels of rHA, granulocyte-macrophage colony-stimulating factor, and recombinant human transferrin.
authors.affiliation: Novozymes Biopharma UK Ltd., Castle Court, 59 Castle Boulevard, Nottingham NG7 1FD, United Kingdom. tpay@novozymes.com
authors.forename: T
authors.initials: T
authors.lastname: Payne
authors.forename: C
authors.initials: C
authors.lastname: Finnis
authors.forename: L R
authors.initials: LR
authors.lastname: Evans
authors.forename: D J
authors.initials: DJ
authors.lastname: Mead
authors.forename: S V
authors.initials: SV
authors.lastname: Avery
authors.forename: D B
authors.initials: DB
authors.lastname: Archer
authors.forename: D
authors.initials: D
authors.lastname: Sleep
mesh_terms: D000418:Albumins / Q000235:genetics / Q000378:metabolism; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000378:metabolism; D006801:Humans; D018832:Molecular Chaperones / Q000096:biosynthesis* / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis / Q000235:genetics; D014168:Transferrin / Q000235:genetics / Q000378:metabolism
chemical_list: D000418:Albumins; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D014168:Transferrin; D016178:Granulocyte-Macrophage Colony-Stimulating Factor